Alteration of Microstructure in Biopolymeric Hydrogels via Compositional Modification of Resilin-Like Polypeptides
Alteration of Microstructure in Biopolymeric Hydrogels via Compositional Modification of Resilin-Like Polypeptides
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DOI：
10.1021/acsbiomaterials.0c01543
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发表时间：
2021-01-19
影响因子：
5.8
通讯作者：
Kiick, Kristi L.
中科院分区：
文献类型：
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作者：
Garcia, Cristobal Garcia;Patkar, Sai S.;Kiick, Kristi L.
已知水凝胶支架中的异质性会影响载有细胞的材料构建体中细胞的性能，并且我们已经采用节枝弹性蛋白样多肽（RLP）的相分离作为产生此类材料的手段。在这里，我们研究了节枝弹性蛋白样多肽（RLP）的组成特征，进一步使我们能够控制它们的液-液相分离（LLPS）以及这种控制如何影响微结构水凝胶的形成。在硫酸铵溶液中的RLP的相分离的评估提供了洞察RLP的解决方案，和原子模拟，沿着与二维核奥弗豪泽效应光谱（NOESY）和相关光谱（COSY）H-1 NMR序列依赖的LLPS，建议特定的氨基酸相互作用，可能介导这种相行为。RLP的丙烯酰胺官能化使得它们能够光交联成水凝胶，并且还增强多肽的相分离。加热-冷却方案促进稳定乳液的形成，所述稳定乳液产生具有可调流变性质的不同微结构形态。这些发现提供了用于选择具有相行为的RLP组合物的方法，所述相行为可以容易地随温度的差异进行调节，以在可用于生物应用的方案中控制所得到的非均相水凝胶的形态和机械行为。
Heterogeneities in hydrogel scaffolds are known to impact the performance of cells in cell-laden materials constructs, and we have employed the phase separation of resilin-like polypeptides (RLPs) as a means to generate such materials. Here, we study the compositional features of resilin-like polypeptides (RLPs) that further enable our control of their liquid-liquid phase separation (LLPS) and how such control impacts the formation of microstructured hydrogels. The evaluation of the phase separation of RLPs in solutions of ammonium sulfate offers insights into the sequence-dependent LLPS of the RLP solutions, and atomistic simulations, along with 2D-nuclear Overhauser effect spectroscopy (NOESY) and correlated spectroscopy (COSY) H-1 NMR, suggest specific amino acid interactions that may mediate this phase behavior. The acrylamide functionalization of RLPs enables their photo-cross-linking into hydrogels and also enhances the phase separation of the polypeptides. A heating-cooling protocol promotes the formation of stable emulsions that yield different microstructured morphologies with tunable rheological properties. These findings offer approaches for choosing RLP compositions with phase behaviors that can be easily tuned with differences in temperature to control the resulting morphology and mechanical behavior of the heterogeneous hydrogels in regimes useful for biological applications.